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Short report
Aleta R. Steevens
University of Minnesota
Corresponding Author
ORCiD: https://orcid.org/0000-0001-7104-7816
License:
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This research is the first to produce induced pluripotent stem cell-derived inner ear sensory neurons in the Neurog1 +/- heterozygote mouse using blastocyst complementation. Additionally, this approach corrected non-sensory deficits associated with Neurog1 heterozygosity, indicating that complementation is specific to endogenous Neurog1 function. This work validates the use of blastocyst complementation as a tool to create novel insight into the function of developmental genes and highlights blastocyst complementation as a potential platform for generating chimeric inner ear cell types that can be transplanted into damaged inner ears to improve hearing.
Keywords
Inner ear, Spiral Ganglion Neurons, Neurogenin1, Blastocyst Complementation, Stem Cells, Regenerative Medicine
Figure 1
Figure 2
Figure 3
Figure 4
This is a list of supplementary files associated with this preprint. Click to download.
- hetwtcomparisonimovie.mp4
Supplemental Movie 1. An 8 second video showing segmented and three-dimensionally reconstructed Neurog1+/+ and Neurog1+/- inner ears rotating horizontally, to highlight the morphological differences observed between wild type and non-complemented Neurog1+/- inner ears.
- SupplementalMaterials.pdf
Supplemental Fig 1. Representative gel images of genotyping for a litter from the Neurog1 colony. Top gel is the mutant genotyping, and the bottom gel is the wild type genotyping for the same samples. Wild type (lane 1): 198 bp. Heterozygote (lanes 2-4,6-13): 198 bp and ~ 500 bp. Mutant (lane 5): ~ 500 bp. Heterozygote positive controls (lanes 11-13) were always run in triplicate due to occasional variability in their result. All genotyping was run 3 times for each sample to confirm genotype. Supplemental Fig 2. Representative examples of GFP-expressing induced pluripotent stem cell (iPSC) colonies. (A) GFP expression. (B) Phase Contrast. Note that all of the iPSCs are GFP-labeled. Scale Bar: 100 µm. Supplemental Fig. 3. 2D optical section through a wild type cochlea using sTSLIM. Bracket indicates the sensory cells of the organ of Corti. Note the high degree of resolution throughout the image, in particular the cell bodies in the SGN. Scale Bar: 100 µm. oC: organ of Corti; SGN: Spiral ganglion neurons
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Peer Review Timeline
CURRENT STATUS: Published
View the published article at Stem Cell Research & Therapy.
Version 2
Posted 28 Dec, 2020
No community comments so far
Editorial decision: Accept
On 19 Jan, 2021
Review #1 received
Received 05 Jan, 2021
Review #2 received
Received 31 Dec, 2020
Reviewer #2 agreed
On 27 Dec, 2020
Reviewer #1 agreed
On 25 Dec, 2020
Reviewers invited
Invitations sent on 24 Dec, 2020
Editor assigned
On 08 Dec, 2020
Submission checks complete
On 08 Dec, 2020
Editor invited
On 08 Dec, 2020
No comments provided
Editorial decision: Major Revision
On 15 Nov, 2020
Review #1 received
Received 10 Nov, 2020
Review #2 received
Received 26 Sep, 2020
Reviewer #2 agreed
On 16 Sep, 2020
Reviewer #1 agreed
On 08 Sep, 2020
Reviewers invited
Invitations sent on 05 Sep, 2020
Editor assigned
On 12 Aug, 2020
Submission checks complete
On 11 Aug, 2020
Editor invited
On 11 Aug, 2020
First submitted
On 10 Aug, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Stem Cell & Developmental Cell Biology
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See the published version of this preprint at Stem Cell Research & Therapy.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Short report
Aleta R. Steevens
University of Minnesota
Corresponding Author
ORCiD: https://orcid.org/0000-0001-7104-7816
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Stem Cell Research & Therapy.
Version 2
Posted 28 Dec, 2020
No community comments so far
Editorial decision: Accept
On 19 Jan, 2021
Review #1 received
Received 05 Jan, 2021
Review #2 received
Received 31 Dec, 2020
Reviewer #2 agreed
On 27 Dec, 2020
Reviewer #1 agreed
On 25 Dec, 2020
Reviewers invited
Invitations sent on 24 Dec, 2020
Editor assigned
On 08 Dec, 2020
Submission checks complete
On 08 Dec, 2020
Editor invited
On 08 Dec, 2020
No comments provided
Editorial decision: Major Revision
On 15 Nov, 2020
Review #1 received
Received 10 Nov, 2020
Review #2 received
Received 26 Sep, 2020
Reviewer #2 agreed
On 16 Sep, 2020
Reviewer #1 agreed
On 08 Sep, 2020
Reviewers invited
Invitations sent on 05 Sep, 2020
Editor assigned
On 12 Aug, 2020
Submission checks complete
On 11 Aug, 2020
Editor invited
On 11 Aug, 2020
First submitted
On 10 Aug, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Stem Cell & Developmental Cell Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Stem Cell Research & Therapy.
This research is the first to produce induced pluripotent stem cell-derived inner ear sensory neurons in the Neurog1 +/- heterozygote mouse using blastocyst complementation. Additionally, this approach corrected non-sensory deficits associated with Neurog1 heterozygosity, indicating that complementation is specific to endogenous Neurog1 function. This work validates the use of blastocyst complementation as a tool to create novel insight into the function of developmental genes and highlights blastocyst complementation as a potential platform for generating chimeric inner ear cell types that can be transplanted into damaged inner ears to improve hearing.
Figure 1
Figure 2
Figure 3
Figure 4
This is a list of supplementary files associated with this preprint. Click to download.
- hetwtcomparisonimovie.mp4
Supplemental Movie 1. An 8 second video showing segmented and three-dimensionally reconstructed Neurog1+/+ and Neurog1+/- inner ears rotating horizontally, to highlight the morphological differences observed between wild type and non-complemented Neurog1+/- inner ears.
- SupplementalMaterials.pdf
Supplemental Fig 1. Representative gel images of genotyping for a litter from the Neurog1 colony. Top gel is the mutant genotyping, and the bottom gel is the wild type genotyping for the same samples. Wild type (lane 1): 198 bp. Heterozygote (lanes 2-4,6-13): 198 bp and ~ 500 bp. Mutant (lane 5): ~ 500 bp. Heterozygote positive controls (lanes 11-13) were always run in triplicate due to occasional variability in their result. All genotyping was run 3 times for each sample to confirm genotype. Supplemental Fig 2. Representative examples of GFP-expressing induced pluripotent stem cell (iPSC) colonies. (A) GFP expression. (B) Phase Contrast. Note that all of the iPSCs are GFP-labeled. Scale Bar: 100 µm. Supplemental Fig. 3. 2D optical section through a wild type cochlea using sTSLIM. Bracket indicates the sensory cells of the organ of Corti. Note the high degree of resolution throughout the image, in particular the cell bodies in the SGN. Scale Bar: 100 µm. oC: organ of Corti; SGN: Spiral ganglion neurons
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